The E3 ubiquitin ligase LNX1p80 promotes the removal of claudins from tight junctions in MDCK cells

Takahashi, Shouji; Iwamoto, Noriko; Sasaki, Hiroyuki; Ohashi, Masato; Oda, Yusuke; Tsukita, Shöichiro; Furuse, Mikio

doi:10.1242/jcs.040055

Cited by 94 publications

(114 citation statements)

References 37 publications

(56 reference statements)

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“…The constitutive turnover of claudin-2 is thought be mediated by an E3 ubiquitin ligase that targets claudins for subsequent lysosomal degradation (16). However, the signaling pathways that regulate claudin-2 turnover have not been established.…”

Section: Matriptase Regulates Claudin-2 Turnover Via Activation Of Atmentioning

confidence: 99%

Membrane-anchored serine protease matriptase regulates epithelial barrier formation and permeability in the intestine

Buzza

Netzel‐Arnett

Shea‐Donohue³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

154

192

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The intestinal epithelium serves as a major protective barrier between the mammalian host and the external environment. Here we show that the transmembrane serine protease matriptase plays a pivotol role in the formation and integrity of the intestinal epithelial barrier. St14 hypomorphic mice, which have a 100-fold reduction in intestinal matriptase mRNA levels, display a 35% reduction in intestinal transepithelial electrical resistance (TEER). Matriptase is expressed during intestinal epithelial differentiation and colocalizes with E-cadherin to apical junctional complexes (AJC) in differentiated polarized Caco-2 monolayers. Inhibition of matriptase activity using a specific peptide inhibitor or by knockdown of matriptase by siRNA disrupts the development of TEER in barrierforming Caco-2 monolayers and increases paracellular permeability to macromolecular FITC-dextran. Loss of matriptase was associated with enhanced expression and incorporation of the permeabilityassociated, "leaky" tight junction protein claudin-2 at intercellular junctions. Knockdown of claudin-2 enhanced the development of TEER in matriptase-silenced Caco-2 monolayers, suggesting that the reduced barrier integrity was caused, at least in part, by an inability to regulate claudin-2 expression and incorporation into junctions. We find that matriptase enhances the rate of claudin-2 protein turnover, and that this is mediated indirectly through an atypical PKCζ-dependent signaling pathway. These results support a key role for matriptase in regulating intestinal epithelial barrier competence, and suggest an intriguing link between pericellular serine protease activity and tight junction assembly in polarized epithelia.claudin-2 | intestinal barrier | St14 | type II transmembrane serine protease | tight junction

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Section: Matriptase Regulates Claudin-2 Turnover Via Activation Of Atmentioning

confidence: 99%

Membrane-anchored serine protease matriptase regulates epithelial barrier formation and permeability in the intestine

Buzza

Netzel‐Arnett

Shea‐Donohue³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

154

192

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“…Occludin and ZO-1 are two well known markers of AJC tight junctions, contributing to tight junction stabilization and optimal barrier function (48,49). As shown in Fig.…”

Section: Pkd3 Overexpression Abolishes Claudin-1 and Claudin-4 Localimentioning

confidence: 99%

Protein Kinase D Promotes Airway Epithelial Barrier Dysfunction and Permeability through Down-regulation of Claudin-1

Gan

Wang

Qin

et al. 2013

Journal of Biological Chemistry

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Background: Claudin-1 is a key component of epithelial tight junctions. Results: PKD, especially PKD3, suppresses claudin-1 expression and promotes airway epithelial barrier dysfunction and permeability. Conclusion: PKD is a negative regulator of claudin-1 and airway epithelial barrier integrity. Significant: Our findings offer new insights into the regulation of airway epithelial barrier integrity and a novel therapeutic target for barrier dysfunction-associated airway and lung diseases.

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“…1C) (Dho et al, 1998;Rice et al, 2001). However, in contrast to LNX2, the function of LNX1p80 as E3 ligase has been partially explored (Kansaku et al, 2006;Takahashi et al, 2009). For these reasons, we decided to test whether LNX1 is also a CD8a binding protein.…”

Section: Cd8a Binds the Lnx Proteins Lnx2 And Lnx1p80mentioning

confidence: 99%

“…Next, we examined whether LNX1p80 was able to ubiquitylate CD8a, as previously described for claudin (Takahashi et al, 2009), and whether also LNX2 was endowed of ubiquitylation activity. For this purpose, we coexpressed CD8a, HA-tagged ubiquitin, and GFP, GFP-LNX1p80 or GFP-LNX2 in HEK293 cells (Fig.…”

Section: Lnx1p80 and Lnx2 Promote Ubiquitylation Of Cd8amentioning

confidence: 99%

Ligand of Numb proteins LNX1p80 and LNX2 interact with the human glycoprotein CD8α and promote its ubiquitylation and endocytosis

D’Agostino

Tornillo

Caporaso

et al. 2011

Journal of Cell Science

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SummaryE3 ubiquitin ligases give specificity to the ubiquitylation process by selectively binding substrates. Recently, their function has emerged as a crucial modulator of T-cell tolerance and immunity. However, substrates, partners and mechanism of action for most E3 ligases remain largely unknown. In this study, we identified the human T-cell co-receptor CD8 a-chain as binding partner of the ligand of Numb proteins X1 (LNX1p80 isoform) and X2 (LNX2). Both LNX mRNAs were found expressed in T cells purified from human blood, and both proteins interacted with CD8a in human HPB-ALL T cells. By using an in vitro assay and a heterologous expression system we showed that the interaction is mediated by the PDZ (PSD95-DlgA-ZO-1) domains of LNX proteins and the cytosolic C-terminal valine motif of CD8a. Moreover, CD8a redistributed LNX1 or LNX2 from the cytosol to the plasma membrane, whereas, remarkably, LNX1 or LNX2 promoted CD8a ubiquitylation, downregulation from the plasma membrane, transport to the lysosomes, and degradation. Our findings highlight the function of LNX proteins as E3 ligases and suggest a mechanism of regulation for CD8a localization at the plasma membrane by ubiquitylation and endocytosis.

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The E3 ubiquitin ligase LNX1p80 promotes the removal of claudins from tight junctions in MDCK cells

Cited by 94 publications

References 37 publications

Membrane-anchored serine protease matriptase regulates epithelial barrier formation and permeability in the intestine

Membrane-anchored serine protease matriptase regulates epithelial barrier formation and permeability in the intestine

Protein Kinase D Promotes Airway Epithelial Barrier Dysfunction and Permeability through Down-regulation of Claudin-1

Ligand of Numb proteins LNX1p80 and LNX2 interact with the human glycoprotein CD8α and promote its ubiquitylation and endocytosis

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